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The role of human papillomavirus DNA methylation in cervical lesion progression.January 2011 (has links)
Fung, Man See Joyce. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 111-120). / Abstracts in English and Chinese. / Table of Contents / Acknowledgements --- p.I / Abstract --- p.II / 論文摘要 --- p.VII / Table of Contents --- p.X / List of Figures --- p.XIV / List of Tables --- p.XVI / Abbreviations --- p.XVII / Chapter Chapter 1 - --- Introduction --- p.l / Chapter 1.1 --- Biology of HPV --- p.2 / Chapter 1.1.1 --- History --- p.2 / Chapter 1.1.2 --- Classification --- p.2 / Chapter 1.1.3 --- Genome structure --- p.3 / Chapter 1.2 --- HPV and cervical cancer --- p.8 / Chapter 1.2.1 --- Classification of cervical lesions --- p.8 / Chapter 1.2.2 --- Natural history of development of cervical cancer --- p.9 / Chapter 1.2.3 --- Risk factors --- p.11 / Chapter 1.3 --- Prevention of cervical cancer --- p.12 / Chapter 1.3.1 --- Vaccination --- p.12 / Chapter 1.3.2 --- Screening --- p.12 / Chapter 1.3.2.1 --- Pap test --- p.12 / Chapter 1.3.2.2 --- HPV DNA test --- p.13 / Chapter 1.3.2.3 --- Methylation pattern as a novel marker --- p.13 / Chapter 1.4 --- Biology of Methylation --- p.14 / Chapter 1.4.1 --- Definition --- p.14 / Chapter 1.4.2 --- Silencing effect --- p.18 / Chapter 1.4.3 --- Roles in normal development --- p.20 / Chapter 1.5 --- Methylation and human diseases --- p.20 / Chapter 1.5.1 --- Genetic diseases --- p.20 / Chapter 1.5.2 --- Cancers --- p.21 / Chapter 1.5.3 --- Methylation and oncogenic viruses --- p.23 / Chapter 1.5.4 --- Potential of methylation pattern as a novel biomarker of cancer --- p.24 / Chapter 1.5.5 --- Epigenetic therapy --- p.25 / Chapter 1.6 --- Methylation and HPV --- p.25 / Chapter 1.6.1 --- History --- p.25 / Chapter 1.6.2 --- Potential roles in transcription regulation of HPV --- p.26 / Chapter 1.6.3 --- Viral gene methylation --- p.27 / Chapter Chapter 2 - --- "Hypotheses, Objectives and Study Design" --- p.28 / Chapter 2.1 --- Hypotheses --- p.29 / Chapter 2.2 --- Objectives --- p.30 / Chapter 2.3 --- Study Design --- p.30 / Chapter Chapter 3 - --- Materials and Methods --- p.34 / Chapter 3.1 --- Work flow --- p.35 / Chapter 3.2 --- Study subjects --- p.37 / Chapter 3.2.1 --- Invasive cervical cancer group --- p.37 / Chapter 3.2.2 --- Low-grade group --- p.37 / Chapter 3.2.3 --- Cell lines --- p.38 / Chapter 3.3 --- DNA extraction --- p.38 / Chapter 3.4 --- HPV genotyping --- p.39 / Chapter 3.5 --- PCR of HPV16 LCR --- p.39 / Chapter 3.6 --- Sequencing of HPV 16 LCR --- p.42 / Chapter 3.6.1 --- Purification of PCR products --- p.42 / Chapter 3.6.2 --- Cycle sequencing reaction --- p.42 / Chapter 3.6.3 --- Purification of cycle sequencing products --- p.43 / Chapter 3.6.4 --- Sequencer and data analysis --- p.43 / Chapter 3.7 --- Bisulfite modification --- p.43 / Chapter 3.8 --- PCR of bisulfite modified LCR --- p.45 / Chapter 3.9 --- Cloning --- p.48 / Chapter 3.9.1 --- Ligation --- p.48 / Chapter 3.9.2 --- Transformation --- p.48 / Chapter 3.9.3 --- Colony PCR --- p.49 / Chapter 3.10 --- Sequencing of clones --- p.51 / Chapter 3.10.1 --- Purification of PCR products --- p.51 / Chapter 3.10.2 --- Cycle sequencing reaction --- p.51 / Chapter 3.10.3 --- Purification of cycle sequencing products --- p.52 / Chapter 3.10.4 --- Sequencer and data analysis --- p.52 / Chapter 3.11 --- Statistical methods --- p.52 / Chapter Chapter 4 - --- Results --- p.54 / Chapter 4.1 --- Sample selection --- p.55 / Chapter 4.2 --- HPV16 LCR PCR and sequencing --- p.57 / Chapter 4.3 --- Methylation patterns --- p.61 / Chapter 4.3.1 --- Cell lines --- p.61 / Chapter 4.3.2 --- Cancer group --- p.63 / Chapter 4.3.2.1 --- Overview --- p.63 / Chapter 4.3.2.2 --- Methylation pattern of the cancer samples --- p.66 / Chapter 4.3.2.3 --- Methylation pattern of the promoter region --- p.74 / Chapter 4.3.3 --- Low-grade group --- p.76 / Chapter 4.3.3.1 --- Overview --- p.76 / Chapter 4.3.3.2 --- Methylation pattern of the low-grade samples --- p.79 / Chapter 4.3.4 --- Comparison of the methylation patterns of the cancer samples and the low-grade samples --- p.84 / Chapter Chapter 5 - --- Discussion --- p.95 / Chapter 5.1 --- Sequence variations of HPV 16 LCR --- p.96 / Chapter 5.2 --- Methylation patterns of CaSki and SiHa cell lines --- p.98 / Chapter 5.3 --- Methylation pattern of the cancer samples --- p.99 / Chapter 5.4 --- Methylation pattern of the low-grade samples --- p.100 / Chapter 5.5 --- Comparison of methylation patterns of the cancer samples and the low-grade samples --- p.101 / Chapter 5.5.1 --- Promoter region in 3' LCR --- p.102 / Chapter 5.5.1.1 --- SP1 binding site --- p.102 / Chapter 5.5.1.2 --- E2BS3 and E2BS4 --- p.103 / Chapter 5.5.2 --- Silencer region --- p.104 / Chapter 5.5.3 --- Enhancer region in central LCR --- p.105 / Chapter 5.5.4 --- CpG sites within 5' LCR --- p.106 / Chapter 5.6 --- Role of methylation in HPV 16 --- p.107 / Chapter 5.7 --- Potential as novel biomarker --- p.108 / Chapter 5.8 --- Conclusions --- p.109 / References --- p.111 / Appendix A
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Prevalence and intra-type variation of human papillomavirus (HPV) infection in cervical cancers: a nationwide perspective of China.January 2001 (has links)
Li Chun-bong. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 147-169). / Abstracts in English and Chinese. / Abstract --- p.i / Declaration --- p.vi / Acknowledgments --- p.vii / Table of Contents --- p.xi / List of Figures --- p.xii / List of Tables --- p.xvi / Abbreviations --- p.xvii / Chapter CHAPTER 1 --- INTRODUCTION AND LITERATURE REVIEWS / Chapter 1.1 --- Introduction --- p.1 / Chapter 1.2 --- Carcinoma of the cervix --- p.6 / Chapter 1.2.1 --- Squamous carcinoma --- p.6 / Chapter 1.2.2 --- Adenosquamous carcinoma --- p.7 / Chapter 1.2.3 --- Adenocarcinoma --- p.8 / Chapter 1.3 --- Molecular biology of Human papillomavirus --- p.9 / Chapter 1.3.1 --- Genome structure and organization of HPV --- p.9 / Chapter 1.3.2 --- Expression of papillomavirus genes --- p.11 / Chapter 1.3.3 --- Taxonomy of HPV --- p.20 / Chapter 1.4 --- Diagnostic techniques in HPV detection --- p.23 / Chapter 1.4.1 --- Southern blot analysis --- p.23 / Chapter 1.4.2 --- Dot blot analysis --- p.25 / Chapter 1.4.3 --- In situ hybridization --- p.26 / Chapter 1.4.4 --- Hybird Capture System --- p.28 / Chapter 1.4.5 --- Polymerase Chain Reaction --- p.30 / Chapter 1.5 --- Human papillomavirus in cervical carcinoma --- p.33 / Chapter 1.5.1 --- Prevalence --- p.33 / Chapter 1.5.2 --- Transmission --- p.37 / Chapter 1.5.3 --- Risk Factors --- p.39 / Chapter CHAPTER2 --- MATERIALS AND METHODS / Chapter 2.1 --- Materials --- p.44 / Chapter 2.1.1 --- Chemicals and regents --- p.44 / Chapter 2.1.2 --- Specimens collection --- p.48 / Chapter 2.2 --- Methods --- p.49 / Chapter 2.2.1 --- Summary of methodology --- p.50 / Chapter 2.2.2 --- DNA extraction from fresh and paraffin embedded tissues --- p.51 / Chapter 2.2.3 --- Polymerase Chain Reaction using HPV Consensus Primer MY09/11 --- p.55 / Chapter 2.2.3.1 --- Template for PCR --- p.55 / Chapter 2.2.3.2 --- PCR amplification --- p.55 / Chapter 2.2.3.3 --- PCR product analysis --- p.56 / Chapter 2.2.4 --- DNA sequencing --- p.57 / Chapter 2.2.4.1 --- DNA sequencing reaction for ALFexpress DNA automatic sequencing --- p.57 / Chapter 2.2.4.2 --- ABI comparative PCR sequencing --- p.59 / Chapter 2.2.4.3 --- DNA sequence analysis --- p.60 / Chapter 2.2.5 --- Restriction Fragment Length Polymorphism --- p.61 / Chapter 2.2.5.1 --- Template preparation --- p.61 / Chapter 2.2.5.2 --- Restriction enzyme digestion --- p.62 / Chapter 2.2.5.3 --- Agarose gel electrophoresis analysis --- p.62 / Chapter 2.2.6 --- HPV Type Specific PCR --- p.63 / Chapter 2.2.6.1 --- Preparation of positive control DNA --- p.63 / Chapter 2.2.6.2 --- Preparation of HPV 52 and HPV 58 type specific PCR --- p.63 / Chapter 2.2.6.3 --- PCR primer design --- p.66 / Chapter 2.2.6.4 --- PCR amplification --- p.68 / Chapter 2.2.7 --- Polymerase Chain Reaction using HPV Consensus Primer GP5+/6+ --- p.71 / Chapter 2.2.7.1 --- Template for PCR --- p.71 / Chapter 2.2.7.2 --- PCR amplification --- p.71 / Chapter 2.2.7.3 --- PCR product analysis --- p.72 / Chapter 2.2.8 --- Statistical analysis --- p.72 / Chapter CHAPTER3 --- RESULTS / Chapter 3.1 --- Histology review of tumor specimens --- p.73 / Chapter 3.2 --- Polymerase chain reaction of HPV consensus primer MY09/11 --- p.76 / Chapter 3.3 --- DNA sequencing reaction --- p.81 / Chapter 3.4 --- Restriction fragment length polymorphism --- p.86 / Chapter 3.5 --- HPV type specific polymerase chain reaction --- p.90 / Chapter 3.6 --- Polymerase chain reaction of HPV consensus primer GP5+/6+ --- p.109 / Chapter 3.7 --- "Correlations of HPV prevalence, geographical variation, histology and age of the cervical cancer patients" --- p.112 / Chapter CHAPTER4 --- DISCUSSION / Chapter 4.1 --- Prevalence of HPV infection in cervical cancer in China --- p.118 / Chapter 4.2 --- DNA extraction and detection methods --- p.131 / Chapter 4.3 --- Intratype variation of HPV --- p.141 / Chapter CHAPTER5 --- CONCLUSION AND FUTURE PERSPECTIVE --- p.143 / REFERENCES --- p.147 / RELEVANT PUBLICATIONS --- p.170
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HIV-1, HIV-2, and dual infection with HIV-1 and HIV-2 are associated with increased risk for human papillomavirus (HPV) and high grade squamous intraepithelial lesions (HSIL) in Senegal, West Africa /Hawes, Stephen Edward. January 2001 (has links)
Thesis (Ph. D.)--University of Washington, 2001. / Vita. Includes bibliographical references (leaves 71-78).
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The role of human papillomavirus in adenocarcinoma of the uterine cervix /Andersson, Sonia, January 2002 (has links)
Diss. (sammanfattning) Stockholm : Karol. inst., 2002. / Härtill 5 uppsatser.
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Longitudinal studies of human papillomavirus infection : with special reference to screening for cervical cancer and treatment of CIN /Elfgren, Kristina, January 1900 (has links)
Diss. (sammanfattning) Stockholm : Karol. inst., 2003. / Härtill 5 uppsatser.
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Improved human papillomavirus DNA typing methods and biology of cervical cancer /Zheng, Biying, January 2006 (has links)
Diss. (sammanfattning) Stockholm : Karol. inst., 2006. / Härtill 5 uppsatser.
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Impact of immunosuppression on the incidence and clearance of human papillomavirus in HIV-infected women in AlabamaBhatta, Madhav P. January 2007 (has links) (PDF)
Thesis (Ph. D.)--University of Alabama at Birmingham, 2007. / Title from first page of PDF file (viewed Oct. 13, 2008). Includes bibliographical references.
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Genetic variation in immune regulatory cytokine genes, cigarette smoking, and human papillomavirus-associated cancer risk /Hussain, Shehnaz Khursheed. January 2006 (has links)
Thesis (Ph. D.)--University of Washington, 2006. / Vita. Includes bibliographical references (leaves 67-80).
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Genital HPV infection and E7 mRNA viral load : incidence, risk factors, and relations to genital neoplasias /Winer, Rachel L. January 2005 (has links)
Thesis (Ph. D.)--University of Washington, 2005. / Vita. Includes bibliographical references (leaves 72-106).
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Revisão sistemática sobre os estudos de prevalência de infecção do colo do útero pelo papilomavírus humano (HPV) no Brasil / Systematic review of prevalence studies of infection of the cervix by human papillomavirus (HPV) in BrazilAndréia Rodrigues Gonçalves Ayres 10 September 2009 (has links)
O câncer do colo do útero é responsável por 7% do total dos óbitos por câncer entre a população feminina brasileira e tem uma incidência estimada de 20/100 mil para todo o país.
Evidências científicas comprovam que o papilomavírus humano (HPV) é causa necessária para a ocorrência deste tipo de câncer. Ações de prevenção e controle recomendadas têm se
baseado no conhecimento sobre a epidemiologia da doença. Os estudos realizados no Brasil sobre a prevalência da infecção por HPV disponíveis na literatura têm características variadas que ainda não foram analisadas em conjunto e de modo sistematizado. O objetivo deste estudo foi realizar uma revisão sistemática dos artigos sobre prevalência do HPV em mulheres
brasileiras considerando as prevalências globais e entre aquelas com exame citológico cervical normal. Foram selecionados todos os artigos após busca nas bases de dados Medline e BVS, tomando-se como termos human papillomavirus, HPV, prevalence Brazil. Entre 1989 e 2008, foram selecionados 155 artigos, sendo 133 nas bases de dados e 22 referências secundárias. Após leitura de título e resumo, 82 artigos foram incluídos, e a seguir submetidos à leitura integral dos textos, sendo enfim selecionados 14 artigos, os quais representaram estudos de quatro grandes regiões brasileiras (Sudeste 43,0%, Sul 21,4%, Nordeste 21,4% e Norte 7,1%). Em sua maioria (64,5%), trata-se de artigos que relatam desenho transversal. Com referência ao método de identificação do HPV nas mulheres, em oito (57,1%) artigos, há relato do emprego de PCR para tipagem do HPV e, em sete (50,0%) artigos, houve emprego de HC para detecção do HPV. As amostras variaram de 49 a 2329 mulheres. A prevalência global de infecção do colo do útero pelo HPV variou entre 13,7 e 54,3%, e para as mulheres com citologia normal, a prevalência de infecção pelo HPV no colo
do útero varia entre 10 e 24,5%. Os resultados obtidos permitiram criar um panorama das prevalências e da distribuição da infecção pelo HPV e principais tipos em mulheres com citologia cervical normal e assim contribuir para a compreensão da distribuição da infecção pelo HPV no país, auxiliando na orientação de outros estudos bem como de políticas voltadas
para a saúde da mulher e prevenção do câncer do colo do útero. / Cervical cancer causes 7% of cancer deaths among Brazilian female population and has an estimated incidence rate of 20/100 thousand for the country. Scientific evidences proof
that human papillomavirus (HPV) is necessary cause to this cancer. Control and prevention actions are recommended with the knowledge about the disease epidemiology. Brazilian studies about HPV prevalence in literature have different characteristics and wait to be analyzed in a set and in systematized manner. The aim of this study was perform a systematic review of the papers about HPV prevalence in Brazilian among Brazilian women, considering global prevalence and those with normal cytology results. All papers were selected after search in Medline and BVS databases, using terms human papillomavirus HPV prevalence Brazil NOT HIV NOT pregnant. Between 1989 and 2008, 155 papers were selected (133 in Medline, 22 secondary references). After reading of title and abstract,
82 papers were included, and then submitted to perusal of the texts; at the end, 14 papers were selected, representing studies from four great Brazilian regions (Southeast 43%, South 21,4%, Northeast 21,4% e North 7,1%). In 64,5% papers, the studies were cross-sectional. Concerning the HPV identifying method in women, in eight (57,1%) papers PCR was used to typing HPV and in seven (50%) papers HC was used to HPV detection. The samples range from 49 to 2329 women. HPV global prevalence in cervix range from 13,7 to 54,3%, and for
women with normal cytology results, HPV prevalence in cervix varied from 10 e 24,5%. The results of this study may contribute to the understanding about distribution of HPV infection in country, helping to the guidance of other studies, as well as in the politics focused to womens health and control and prevention of cervical cancer.
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